Characterization of White Shrimp Litopenaeus Vannamei Pattern Recognition Protein Receptor and Its Role Involved in the Immune Initiation.

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基本資料

Project title

Code/計畫編號

NSC98-2313-B019-002-MY3

Translated Name/計畫中文名

白蝦辨識蛋白受體在免疫啟動特性研究

Project Coordinator/計畫主持人

Jiann-Chu Chen

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Aquaculture

Website

https://www.grb.gov.tw/search/planDetail?id=2222709

Year

2011

Start date/計畫起

01-08-2011

Expected Completion/計畫迄

31-07-2012

Bugetid/研究經費

1500千元

ResearchField/研究領域

漁業
生物技術(農)

白蝦為全世界相當重要的養殖蝦種，在養殖過程中，常受到弧菌、黴菌、寄生蟲等的侵襲。蝦缺乏適應性反應，完全依賴先天性免疫，因此研究蝦先天性免疫防禦，瞭解其免疫啟動機制，為當務之急。一旦細菌等異物入侵，進入體腔時，即啟動一連串複雜的先天性免疫反應。參與先天性免疫系統的蛋白質，可以分成幾大類，分別為 1)辨識蛋白 (Pattern recognition proteins, PRPs )包括LGBP、βGBP、PGRP 及lectin 等，2)原酚氧化酵素活化系統相關蛋白質包括prophenoloxidase、peroxinectin、serine proteinase、proteinase inhibitors 如 pacifastin 及α2-macroglobulin，3)參與吞噬作用，反應氧族群產生及抗氧化系統蛋白質如 SOD、peroxidase，catalase 等，4)凝集系統蛋白質包括clottable protein 及transglutaminase。 5)抗菌胜肽如penaeidin，crustin 及ALF 等，6) lysozyme。其中以辨識蛋白扮演啟動先天性免疫反應的重要角色。白蝦Litopenaeus vannamei βGBP 及LGBP 已有選殖研究，但是關於此等辨識蛋白質受體如LGBP 受體integrin 的選殖、特性，LGBP 如何與受體intergin 結合，以及peroxinectin 如何與受體intergin 結合，啟動並促進先天性免疫防禦的研究厥如。因此，本計畫擬在第一年選殖白蝦辨識蛋白LGBP 的受體integrin。第二年研究白蝦 LGBP，integrin 及peroxinectin 在組織的分佈，在不同脫殼期，以及當蝦在受到異物如LPS、 β-1-3-glucan、peptidoglycan、zymosan 及Vibrio alginolyticus 入侵，環境因子變化如水溫、鹽度及低溶氧緊迫下的表現。第三年擬用酵母Pichia pastoris 重組表現系統來生產表現此等LGBP，integrin 與peroxinectin 蛋白質，進而研究此等重組免疫蛋白質功能及結合與啟動特性。 White shrimp Litopenaeus vannamei, tiger shrimp Penaeus monodon, and kuruma shrimp Marsupenaeus japonicus are important penaeid shrimps currently being cultured in the world. Culture of white shrimp L. vannamei has become primary species in Asia, where it accounts 90% of total production in the world. However, shrimp culture has been suffered by viral disease and vibriosis leading to vast economic loss. Shrimp like other invertebrate lacks a true adaptive immune system, and relies instead on innate responses against invading pathogens. Once pathogens enter the hemocoel of the host, they may encounter a complex system of innate immune systems. There are several proteins involved in the innate immune system. They are 1) pattern recognition proteins (PRPs) like LPS and β-1,3-glucan binding protein (LGBP), β-1,3-glucan binding protein (βGBP), peptidoglcan recognition protein (PGRP), and lectin etc, 2) prophenoloxidase activating system proteins like prophenoloxidase, peroxinectin, serine proteinase (prophenoloxidase activating enzyme, ppA), proteinase inhibitors like pacifastin and α2-macroglobulin, 3) phagocytosis, reactive oxygen intermediates (ROIs) and antioxidant protective enzymes like SOD, peroxidase and catalase, 4) coagulation system proteins like clottable protein and transglutaminase, 6) anti-microbial peptides (AMP) like penaeidin, crustin and ALF, and 6) lysozyme. Among these proteins, PRPs play the most important role in the initiation of innate immune system. The LGBP and βGBP have been cloned and characterized from white shrimp L. vannamei. However, none is kown on the cloning of integrin-like protein, and the cross-link between LGBP and integrin as well as the cross-link between integrin and peroxinectin that initiate and enhance the innate system of penaeid shrimp. This proposal is to study cDNA cloning of integrin and its characterization of white shrimp L. vannamei during the first year. This proposal is to study LGBP, intergin, and peroxinectin expression of white shrimp L. vannamei in different tissues, molting stages, and when white shrimp L. vannamei receive LPS, β-1,3-glucan, peptidoglycan, zymosan, V. alginolyticus, and under environmental stress like water temperature, salinity and hypoxia during the second year. This proposal is to study the expression of LGBP and integrin, and peroxinectin using yeast Pichia pastoris expression system to produce the recombinant protein and to study their binding, functions and activities of white shrimp during the third year.

Keyword(s)

白蝦
辨識蛋白
LGBP
integrin-like proteins
peroxinectin
基因選殖
LPS
_x000d_ glucan
peptidoglycan
脫殼期
組織
水溫
鹽度
低溶氧
基因表現
重組蛋白
White shrimp
LGBP
integrin protein
peroxinectin
LPS
β-1
3-glucan
zymosan
_x000d_ peptidoglycan
carrageenan
moulting stages
tissues
temperature
salinity
hypoxia
Gene_x000d_ expression
Recombinant protein
Pichia pastoris

DSpace CRIS

Characterization of White Shrimp Litopenaeus Vannamei Pattern Recognition Protein Receptor and Its Role Involved in the Immune Initiation.

基本資料

Description